Our earlier findings that epithelial ovarian cancer (EOC) is recognized by the host's immune response, and that activation of antitumor immune response is associated with longer survival, revive hopes that immune interventions might offer important therapeutic opportunities in EOC. However, current immune therapy approaches have generally insufficient power to induce rapid and oven/vhelming tumor rejection response. Over the past SPORE funding cycle, Drs Coukos, June and colleagues have launched a clinical immunotherapy program in ovarian cancer, made important advances in understanding mechanisms that disable or attenuate tumor immune attack, and developed tools to counteract these. We have found that tumor-infiltrating lymphocytes recognize autologous tumor and once expanded appropriately ex vivo, they can suppress tumor growth in vivo following adoptive transfer. We propose to test the central hvpothesis that it is now possible for the first time to deliver powerful immunotherapy for ovarian cancer capitalizing on tumor-infiltrating lymphocytes (TIL) optimized for adoptive therapy through advanced culture platforms we developed. A related hypothesis is that it is now possible to break the tumor barriers to immune attack by (a) restoring tumor antigen presentation and tumor recognition by T cells (Signal 1) through rational use of chemotherapy; and (b) restoring costimulatory activation of T cells at the tumor site (Signal 2) through engineering T cells with costimulatory chimeric immunoreceptors (CIRs) redirected against tumor antigens. Specific Aim 1 will translate laboratory findings into adoptive lymphocyte therapy trials. A phase l/ll clinical trial will be conducted to test the hypothesis that appropriate costimulation ex vivo of TIL and host lymphodepletion augment the response to adoptive T cell transfer therapy. Specific Aim 2 proposes to maximize T cell recognition of tumor (Signal 1) through use of chemotherapy. We will test the preclinical effect of combining adoptive TIL therapy with pegylated liposomal doxorubicin (Doxil) and define optimal schedules in order to move to the clinic a combination trial. Specific Aim 3 proposes to maximize in vivo costimulation (Signal 2) through the use of chimeric immunoreceptors. We will test the preclinical efficacy of tumor-derived T cells engineered ex vivo to express costimulatory chimeric immunoreceptors recognizing a common EOC antigen, mesothelin. RELEVANCE (See instructions): Current immune therapy approaches have generally insufficient power to induce rapid and ovenwhelming tumor rejection response. We assert that the advanced techniques for T cell production and pharmacologic and engineering interventions proposed are poised to resolve many of the current limitations in immunotherapy. If we are successful, our work will produce significant therapeutic increments in ovarian cancer and provide important technology for other tumors.